LOCUS       sequence-141764-         866 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  All-in-One plasmid encoding dual U6 promoter-driven sgRNAs and 
            EGFP-coupled Cas9-D10A nickase to enhance efficient and accurate 
            genome editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 866)
  AUTHORS   Chiang TW, le Sage C, Larrieu D, Demir M, Jackson SP
  TITLE     CRISPR-Cas9(D10A) nickase-based genotypic and phenotypic screening 
            to enhance genome editing.
  JOURNAL   Sci Rep. 2016 Apr 15;6:24356. doi: 10.1038/srep24356.
  PUBMED    27079678
REFERENCE   2  (bases 1 to 866)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2016-04-15"; volume: "6:24356. doi"; pages: " 
            10.1038/srep24356"
COMMENT     Sequence Label: AIO-GFP
FEATURES             Location/Qualifiers
     source          1..866
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     203..224
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(267..320)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(336..383)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac
       61 tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
      121 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
      181 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
      241 gtgaacagct cctcgccctt gctcactggg ccaggattct cctcgacgtc accgcatgtt
      301 agcagacttc ctctgccctc tccactgccg aattcctttt tcttttttgc ctggccggcc
      361 tttttcgtgg ccgccggcct tttgtcgcct cccagctgag acaggtcgat ccgtgtctcg
      421 tacaggccgg tgatgctctg gtggatcagg gtggcgtcca gcacctcttt ggtgctggtg
      481 tacctcttcc ggtcgatggt ggtgtcaaag tacttgaagg cggcaggggc tcccagattg
      541 gtcagggtaa acaggtggat gatattctcg gcctgctctc tgatgggctt atcccggtgc
      601 ttgttgtagg cggacagcac tttgtccaga ttagcgtcgg ccaggatcac tctcttggag
      661 aactcgctga tctgctcgat gatctcgtcc aggtagtgct tgtgctgttc cacaaacagc
      721 tgtttctgct cattatcctc gggggagccc ttcagcttct catagtggct ggccaggtac
      781 aggaagttca catatttgga gggcagggcc agttcgtttc ccttctgcag ttcgccggca
      841 gaggccagca ttctcttccg gccgtt
//
